History of Biochemistry Introductory article

Noel G Coley, The Open University, Milton Keynes, UK Article Contents

. Introduction
A history of biochemistry from the eighteenth to the late twentieth century, revealing some . Animal Chemistry
of the problems faced by biochemists seeking chemical explanations for biological . Cell Theory
phenomena and introducing some origins of modern biochemistry. . Biological Oxidation
. Structural Organic Chemistry
. Hormones and Endocrinology
. Vitamins
. Blood Chemistry
. Molecular Biology and the Nucleic Acids

Introduction
istry and molecular energy studies together with physical
Biochemistry aims to explain biological phenomena in techniques such as X-ray diraction, electrophoresis,
chemical terms. The problems are highly complex and until chromatography and labelling with radioisotopes.
the twentieth century progress was slow and unreliable.
During the eighteenth century chemistry was dominated
by the phlogiston theory and the traditional method of
organic analysis by destructive distillation provided no Animal Chemistry
information about elementary composition. Both in
theory and techniques therefore, chemistry was wholly In the 1780s Lavoisier proposed a mechanism for photo-
inadequate to unravel the mysteries of the vital functions. synthesis by which plants take in carbon dioxide and
Yet the importance of fermentation was recognized, release oxygen; he also began to investigate animal
photosynthesis was discovered and studies of animal respiration. With Pierre Laplace he measured the heat
respiration and digestion made useful contributions to evolved by small mammals, and similarities between
early biochemistry. The complex chemical transforma- combustion and respiration led him to locate the produc-
tions occurring at moderate temperatures during the vital tion of animal heat entirely in the lungs. William Prout,
functions led to the belief that in living matter the ordinary better known for his atomic hypothesis, also studied the
laws of chemistry were modied by unknown vital forces. vital processes. He thought that digestion and assimilation
These controlled the special conditions pertaining to living proceeded in stages involving the action of saliva, gastric
matter, but when life was extinguished ordinary chemical juice, pancreatic juice and bile, followed by oxidation in the
forces once more came into play, resulting in decomposi- lungs, where the products of digestion were nally
tion and decay. Despite growing experimental evidence to converted into blood. In 1824 Prout discovered hydro-
the contrary, vitalism persisted among physiologists and chloric acid in gastric juice and he later classied foods as
chemists into the nineteenth century. saccharinous, albuminous and oleaginous (cf. carbohy-
The displacement of phlogiston by oxygen from the late drates, proteins and fats).
eighteenth century greatly facilitated progress. In addition, Michel Euge`ne Chevreul investigated the nature of
Antoine Lavoisiers experimental denition of the chemi- animal fats from about 1811. He showed that the fatty
cal element was accompanied by a more consistent use of acids were analogous to inorganic acids and were capable
the balance and a rational chemical nomenclature. of yielding salts with bases. Separating them using solvents,
Combustion analysis, rst introduced by Joseph Louis he identied each fatty acid by its melting point. The use of
Gay-Lussac in 1811, allowed the empirical formulae of a physical constant as a criterion of purity and means of
organic compounds to be determined, although more identication was an important step in the development of
accurate atomic weights were required for reliable results. organic analysis. His results, published in 1823, showed
Chemists analysed plant and animal substances from the that animal fats, although products of the vital functions,
beginning of the nineteenth century, while physiologists were normal chemical compounds.
and some physicians applied themselves to studies of the Jons Jacob Berzelius and Justus Liebig were among the
vital functions in health and disease. However, the leading nineteenth-century chemists who advanced bio-
development of biochemistry would also require later chemistry. Berzelius analysed animal solids and uids;
nineteenth-century advances in physical and organic Liebig endeavoured to extend Berzeliuss work by creating
chemistry. These included structural organic chemistry, a comprehensive metabolic theory. Beginning with the
electrochemistry, osmosis, colloid chemistry, stereochem- oxygen theory and his own organic analyses, Liebig

ENCYCLOPEDIA OF LIFE SCIENCES 2001, John Wiley & Sons, Ltd. www.els.net 1
 History of Biochemistry

described the assimilation of foods and the oxidation of proteins. In 1878 Willy Kuhne coined the term enzyme for
muscle tissues releasing energy and excretory products like Traubes ferments. Enzyme action had rst been observed
urea and uric acid. For three decades after its publication in in 1833 when Anselme Payen and Jean Francois Persoz
1842, Liebigs Animal Chemistry, or organic chemistry in its isolated a compound (diastase) which converted starch
application to physiology and pathology stimulated chemi- into sugar. Three years later pepsin was extracted from the
cal and physiological research. Unfortunately, Liebig was stomach wall by Schwann. These discoveries preceded
no physiologist and most of those who tried to nd Berzeliuss notion of catalysis (1837) which, he predicted,
supporting evidence for his theories failed. By about 1870 would prove important in plants and animals where
Liebigs theories had been almost entirely superseded, but complex reactions took place at very moderate tempera-
the research they engendered helped to advance biochem- tures. Further investigation of fermentation resulted in a
istry and some nineteenth-century chemists even applied distinction between organized ferments such as yeast and
Liebigs ideas successfully. For example, Henry Bence unorganized ferments including the enzymes, the biolo-
Jones discovered proteins now known to be part of the gical catalysts, but in 1877 Felix Hoppe-Seyler suggested
autoimmune system and Ludwig Thudichum identied that there was no fundamental dierence between them.
some important biochemical substances, including hae- Twenty years later Eduard Buchner fermented a sugar
matoporphyrins in the blood and the main chemical solution with a cell-free extract of yeast. This discovery
constituents of the brain. showed that the contents of the cells, rather than their life
processes, caused fermentation. This led to a theory of
fermentation based on enzyme action and provided the key
to the study of cell chemistry.
Cell Theory Liebigs idea that animals are incapable of biosynthesis
was challenged in the 1850s by Claude Bernards discovery
In 1838 Theodor Schwann, originator of the cell theory in of the glycogenic function of the liver. In 1867 Carl Voit
biology, suggested that fermentation occurred only in living tried to revive Liebigs theory of the direct assimilation of
yeast cells. Liebig refused to accept this and proposed an proteins, but Eduard Puger, Professor of Physiology at
alternative chemical theory of fermentation but Louis Bonn, argued that there were constitutional dierences
Pasteur, opposing Liebigs chemical theory, showed that between food proteins and tissue proteins. In the 1870s he
fermentation depends on the vital functions of living yeast demonstrated the importance of intracellular respiration, a
cells and bacteria. The controversy was resolved in 1872 discovery embraced by Bernard, and suggested a theory of
when Pasteurs work received general recognition. indirect nutrition, whereby animal cells synthesize complex
Close links between plant and animal life have been substances from simpler nutrient molecules derived from
identied. Animal metabolism is ultimately dependent on food. Bernard postulated an internal environment within
the assimilation of plant material and consequently the cells, where chemical degradation and synthesis takes
photosynthesis studies have made important contributions place. According to Bernard all the principles necessary for
to the elucidation of animal metabolism. In 1817 J. Pelletier the maintenance of animal life are released into the blood.
and J. B. Caventou suggested the name chlorophyll for Respiration introduces oxygen, digestion introduces the
the green substance common to all plants. The connection necessary nutrients together with the secretions of the
between chlorophyll and starch in growing plants was various organs. The blood is the carrier of all these
recognized in1862 when Julius von Sachs, a German plant substances and the cells absorb from it only what they
physiologist, suggested that starch in green plants is require to maintain the vital functions occurring inside
produced from carbon dioxide and water. In the early them. The circulation and secretions together ensure,
years of the twentieth century chromatography revealed besides the renewal of the internal environment, the
two forms of chlorophyll and in 1914, Richard Willstatter removal of waste products, and all these changes are
and Arthur Stoll showed that both are esters of dibasic regulated and harmonized by the nervous system, main-
acids with methyl alcohol and phytol, a hitherto unknown taining a steady state (homeostasis).
unsaturated, aliphatic alcohol (C20H39OH). Further in- Bernards theory of the internal environment marks a
vestigation of photosynthesis showed that it involved a watershed in the history of biochemistry. It put an end to
chemical and a photochemical step. The chemical step is theories of direct assimilation, replacing them with the
associated with the formation of a chlorophyllcarbon breakdown of complex food molecules into smaller nutrient
dioxide compound and the photochemical step involves constituents from which new compounds specic to the
the formation of a peroxide which is then decomposed by needs of each cell were synthesized. This theory pregured
an enzyme to yield oxygen, formaldehyde and reconsti- one of the most important discoveries of modern molecular
tuted chlorophyll. biology the coded programme of protein synthesis but
About 1860 Moritz Traube suggested that to understand Bernards ideas of indirect nutrition and the internal
the chemistry of life required a correct theory of fermenta- environment were too advanced for many of his contem-
tion. His ferments were oxygen-rich agents derived from poraries. Much detailed research was needed to establish

2
 History of Biochemistry

them and it was due to British and American scientists vital functions depend on enzyme action. The hydrolytic
including W. M. Bayliss, E. H. Starling, C. S. Sherrington, J. processes catalysed by enzymes break down starch, fats
S. Haldane, J. Barcroft, W. B. Cannon and L. J. Henderson and proteins into simpler molecules (monosaccharides,
that the concept of the internal environment was elaborated fatty acids and amino acids) which then enter complex
from the beginning of the twentieth century. metabolic pathways also controlled by enzymes. During
these processes energy is released, providing animal heat or
to be stored for later use in muscular exertion. One of the
principal aims of twentieth-century biochemistry has been
Biological Oxidation to discover the details of these metabolic pathways.
Considerable progress has been made, though many
It has been recognized from the beginning of the nineteenth details still require further elucidation.
century that energy is released during biological oxidation, In 1907 Walter Fletcher and Frederick Gowland
though the mechanisms by which this was brought about Hopkins published the rst reliable quantitative data on
remained in doubt. When C. F. Schonbein discovered the proportions of lactic acid in muscle tissue. Lactic acid
ozone in 1840 he suggested that the rst step in biological was known to take part in the biochemical transformations
oxidation was the conversion of oxygen to ozone. This idea of carbohydrates, linking them with proteins and fats, but
led to the popular nineteenth-century ozone craze. Later, these two workers now recognized that its role in the
in 1903, A. N. Bach and R. Chodat discovered the enzyme metabolism of animal tissues is pivotal. However, they did
peroxidase in plant cells and it was thought that peroxides not discover that lactic acid holds an intermediate position
caused biological oxidation in a two-stage process. In the between sugar and alcohol in the metabolism of muscle
rst stage the enzyme oxygenase formed peroxides; in the tissue.
second stage peroxidase used these peroxides to oxidize In the early years of the twentieth century Gustav
other organic compounds. This theory dominated ideas on Embden had isolated several intermediate metabolic
biological oxidation until about 1920. Although based on products from muscle tissues, including adenyl phosphoric
plants, it was widely thought that the oxygenase acid, also found in the liver. Embden was the rst to
peroxidase system also accounted for respiration in animal discover and link together all the steps in the conversion of
tissues. It was later found that oxygenase is itself an enzyme glycogen to lactic acid. Between 1919 and 1921 Otto
(catechol oxidase) capable of oxidizing catechol and Meyerhof, recognizing that muscle is the only tissue in
similar compounds to quinones and that peroxidase which it is possible to compare the chemical changes
cannot use these as oxidizing agents. occurring with the work done or heat energy evolved,
From 1920 another theory of biological oxidation was investigated lactic acid formation in muscle tissue as a
proposed, based on the respiratory systems in yeast, measure of work done. This led to the glycogenlactic acid
bacteria and animal tissues. According to this theory, cycle, and made a fundamental contribution to the
oxygen atoms were rst activated by combination with understanding of muscular action. In all but the briefest,
iron atoms in a haemoprotein enzyme. Otto Warburg most intense muscular contractions additional adenosine
called this the respiratory ferment; it would later be triphosphate (ATP), an important energy-carrying coen-
identied as cytochrome oxidase. A rival school considered zyme, is supplied by the chemical reactions of the
that the organic molecules were activated rather than glycolytic, or EmbdenMeyerhofParnas, pathway, par-
oxygen and that most biological oxidations were dehy- ticularly applicable to rapid muscular action. In this, the
drogenations brought about by a new group of enzymes, change from glucose to lactate is coupled with the
the dehydrogenases. These have turned out to be the most formation of ATP from adenosine diphosphate (ADP). If
important factors in biological oxidations; there are now not immediately required, the energy is stored in the
well over 150 known dehydrogenases. By studying these muscle. ATP is created in greater abundance than the
two opposing systems using spectroscopic techniques numbers of carbohydrate, fat or protein molecules
David Keilin followed the course of respiratory changes metabolized, creating more of this energy-rich compound
in the mitochondria of cells. He discovered the oxidative than is required for immediate needs. The excess energy is
cytochromes in 1925 and identied many intermediate stored in muscle tissue as phosphocreatine, a labile
steps in cell respiration. compound that readily absorbs and releases phosphate
In 1906 Arthur Harden and W. J. Young observed that groups.
the addition of a soluble phosphate to a fermenting sugar
Phosphocreatine 1 ADP!creatine 1 ATP
solution caused the rate of fermentation to increase. The
additional carbon dioxide and alcohol formed was
proportional to the quantity of phosphate added and the ATP 2 1 phosphate group!ADP 1 energy
phosphate was converted into a hexosephosphoric acid.
These observations were important for metabolic studies in
the mid-twentieth century, which have shown that most Creatine 1 1 phosphate group!phosphocreatine

3
 History of Biochemistry

The lactate, a waste product, diuses out of the muscle to be cells by a protein enzyme, ATP synthase. The molecular
transported by the blood to the liver where most of it is structure of ATP synthase was partially determined in 1994
converted into glycogen. The rate of oxidation of lactic acid after 12 years of research. In addition to providing fresh
is regulated by the rate of respiration. In prolonged strenuous knowledge about how living things produce energy, it is
exercise lactic acid is formed faster than it can diuse out of thought that this research may throw new light on the
the muscle and this results in muscle fatigue. Glycogen from processes of ageing and the causes of degenerative diseases.
the liver is reversibly converted into glucose and nds its way The large molecules of carbohydrates, fats and proteins are
into the bloodstream, thus completing the cycle. broken down into smaller molecules such as pyruvates, free
In the 1930s Warburg investigated the relation between fatty acids and amino acids, all of which take part in the
the chemical and photochemical steps in photosynthesis mechanisms of the citric acid cycle and thus link the
using the eects of intermittent illumination on the green metabolic transformations of carbohydrates, proteins and
unicellular alga Chlorella. His work led to a study of the fats. Enzymes are now known to control most biochemical
cytochromes. Warburg and Keilin independently exam- processes, from the breakdown of complex molecules like
ined the respiratory function of the cytochromes and carbohydrates and proteins during animal metabolism to
showed that the respiratory chain in the cells was located in the synthesis of macromolecules within the cells. Most
the mitochondria. Alternate oxidation and reduction enzymes have a protein structure and most work with one
could be traced through chains of cytochromes, each or more coenzymes, but certain other complex molecules
reducing the next in line until the last, identied as the not regarded as enzymes, such as messenger RNA, also
respiratory ferment, reacted with oxygen. At each stage the show catalytic properties.
small quantities of energy released are stored in high-
energy bonds such as the phosphate link in ATP. The
whole complex system was summarized in the Krebs cycle,
the metabolic pathways of which have been subjected to Structural Organic Chemistry
minute and intensive research ever since. It also became
clear that other substances besides the cytochromes were Before the details of such cellular mechanisms could be
involved and in 1932 Axel Theorell working with Warburg, fully explored, far more information about the chemical
isolated the rst so-called yellow enzyme composed of a structures of large molecules was needed. Thus, progress in
protein and the nonprotein yellow coenzyme riboavin biochemistry had to await the development of structural
(vitamin B2). Keilin isolated the oxidative enzyme cyto- organic chemistry.
chrome c and proposed a new explanation of the action of In 1815 J. B. Biot found that certain natural oils rotated
the cytochromes involving activated hydrogen as well as the plane of polarized light. The same substance could exist
oxygen. These investigations were carried out on heart- in two forms both having the same empirical formula, yet
muscle preparations, but similar reactions have been capable of rotating the plane of polarized light in opposite
detected in the cells of plants, microorganisms and fungi. directions. The problem facing organic chemists was to
Thus, they represent a common feature shared by all forms correlate optical activity with molecular conguration.
of life. They were essential early stages in the elaboration of The fundamental discovery required to solve this problem
the citric (or tricarboxylic) acid cycle proposed by Sir Hans was Kekules recognition in 1858 that the carbon atom is
Krebs in 1937. In animal cells the enzymes specic to each tetravalent. Using this, J. H. Vant Ho suggested in 1874
step are located in the mitochondria, in plant cells they are that structural formulae must be three-dimensional. A
found in the chloroplasts, and in microorganisms they are compound in which a central carbon atom is attached to
found in the cell walls. four dierent substituents in a tetrahedral structure would
A fuller elaboration of the citric acid cycle came after have two non-superposable forms one of which is the
1940 with the discovery that the three principal food mirror image of the other. Applying this concept Pasteur
constituents, carbohydrates, fats and proteins, all yield a investigated molecular asymmetry in the tartrates by
common product, coenzyme A. This forms citric acid which separating potassium tartrate crystals mechanically. Op-
is then broken down into carbon dioxide and water with the tical activity was later used by Emil Fischer to identify the
liberation of coenzyme A once more. At each intermediate many stereoisomers of the sugars.
step in this metabolic chain, hydrogen atoms are transferred In 1885 Fischer observed that phenylhydrazine
from one carrier to another and small quantities of energy (C6H5.NH.NH2) forms well-dened crystalline compounds
are released. Thus, the citric acid cycle is the source of (osazones) with the sugars and this allowed him to identify
energy refurbishment supporting all the vital processes. the molecular structures of 16 stereoisomers of glucose.
Enzyme reaction rates vary widely and depend on Before 1891 it had been necessary to assume a conguration
activators, or coenzymes, ranging from simple metal ions for each sugar, but from that year onwards Fischer applied
such as Ca2 1 or Mg2 1 to complex organic molecules such himself to the problem of assigning specic congurations to
as vitamins. In 1960 it was found that the coenzyme in each isomeric sugar molecule and by 1896 he had done so for
energy-producing processes, ATP, is itself fabricated in the all the monosaccharides. He retained the straight-chain

4
 History of Biochemistry

formulae devised by H. Kiliani, though later studies by W. N. specic substances which are carried by the blood to other
Haworth and others showed that they contain lactone rings. organs in the body. Towards the end of the nineteenth
Fischer also examined the degradation of sugars by enzymes century a number of discoveries revealed the essential
and from his study of the saccharases he recognized the nature of these secretions for animal health and it became
specicity of enzymes and concluded that the enzyme and its common to connect the lack or excess of specic endocrine
substrate were related as a key to its lock. Later work has secretions with certain diseases. Adrenaline (C9H13O3N)
shown this concept to be true to a degree Fischer himself was discovered in 1894 in experiments on the blood
could hardly have suspected. pressure of a dog. A little later vasopressin, a peptide
Fischers other major contribution to structural organic hormone released from the posterior lobe of the pituitary
chemistry concerned his elucidation of the molecular gland, was discovered. This is also capable of increasing the
structures of the purines, amino acids and proteins. Even blood pressure. Adrenaline was isolated independently in
before he began work on the sugars Fischer had been 1901 by T. B. Aldrich and J. J. Abel in America and shortly
engaged in a study of three purine derivatives, caeine, before by J. Takamine in Japan. In 1902 William Bayliss
theobromine and xanthine, all structurally related to uric and Ernest Starling identied the substance secretin,
acid. Ludwig Medicus had given the correct formula for produced by epithelial cells in the stomach during
uric acid in 1875; Fischer conrmed it by synthesis. digestion. This is released into the bloodstream and in
Between 1882 and 1900 he isolated about 130 purine the pancreas it causes the secretion of pancreatic juice.
derivatives, thus extending Liebig and Friedrich Wohlers Thus, secretin was recognized as a chemical messenger,
1838 studies of the oxidation products of uric acid. Fischer produced in one organ and carried by the bloodstream to
attempted to correlate the molecular structures of these the organ it is intended to stimulate. Starling introduced
compounds with their physiological properties. Later, in the term hormone to describe such chemical messengers
1914, he returned to this work and succeeded in preparing in 1905. However, as secretin is a polypeptide of moderate
the rst synthetic nucleotide, theophylline d-glucose molecular weight, Bayliss and Starling were unable to
phosphoric acid, thus linking his work on sugars with his isolate it, much less determine its molecular structure. The
studies of the purines. This work was a rst step leading to existence of a number of other mammalian hormones was
the study of the nucleo-proteins. Fischer also investigated surmised, but they all proved dicult to isolate and
polypeptides and simple proteins, showing how amino synthesize and only a few had been characterized. It is now
acids are combined in protein molecules. By 1907 he had known that hormones fall into three main classes: steroids,
synthesized a polypeptide containing 18 amino acids. Its peptides and proteins, and amino acid derivatives.
molecular weight was 1213 and he calculated that there Hormone secretion is controlled by the pituitary gland
would be 816 possible optical isomers. situated at the base of the brain. There are three main
In 1877 Traube suggested that enzymes are related to divisions of this gland, although the whole complex organ
proteins, but the isolation of pure enzymes only began in the weighs only about one gram in the human adult. During
1920s. Jackbean urease was the rst enzyme to be crystallized the 1920s work on the pituitary gland led to the isolation of
in a pure state, by J. B. Sumner in 1926. J. H. Northrop, who six dierent protein hormones, four of which were found to
isolated several proteolytic enzymes, crystallized pepsin in stimulate other endocrine glands. Since then other
1930. Both were found to be complex proteins and little hormones have been isolated, several of which control
could be done to determine their structures until special the sexual functions; others govern the rate of growth and
methods were developed. The sequence of amino acids in the chemical or physiological balance in the animal body.
proteins was later found using a method devised by The chemical structures of all have been determined and
Frederick Sanger for determining the order of the 51 amino their chemical syntheses achieved.
acids in the insulin molecule in about 1950. Another group of Adrenaline, a derivative of catechol, was the rst
biological compounds, the hormones, has also caused hormone to be synthesized in 1904; thyroxin, the active
considerable diculty for structural organic chemists in principle of the thyroid gland, rst isolated by E. C.
the twentieth century. Those hormones, which like glucagon Kendall in1914, was synthesized by C. R. Harington and
are polypeptides, have posed problems in the elucidation of G. Barger in 19261927. Thyroxine, stored in the thyroid
their amino acid sequences, but others have been found to be gland as a protein, is hydrolysed by thyrotrophin, another
based on the characteristic ring structure of the sterols hormone produced by the pituitary body. Thus one
containing 17 carbon atoms and 28 hydrogen atoms hormone calls another into action. Thyroxine is mainly
arranged in four rings. concerned with the consumption of oxygen and thus the
metabolism of all the cells and tissues in the body; it
appears to increase the production of a number of
Hormones and Endocrinology enzymes. In 1922 Frederick Banting and Charles Best
identied and prepared the pancreatic hormone, insulin,
In the nineteenth century it was recognized that glands for therapeutic use in diabetes. J. J. Abel obtained crystals
such as the spleen, thymus, thyroid and adrenals secrete of insulin in 1926, but Dorothy Hodgkin using X-ray

5
 History of Biochemistry

diraction elucidated the molecular structure of insulin secretonin. Acetylcholine was rst isolated in 1914; its
only in the 1960s. This work complemented Sangers function in slowing the heartbeat was identied in 1921 by
earlier determination of the complete amino acid sequence Otto Loewi, a German physiologist.
for bovine insulin. Sangers methods opened the way to the In the 1930s it was observed that one hormone can act in
determination of the structures of many other complex opposition to another. Frederic Houssay in Argentina
proteins. He later applied radioisotope labelling using 32P found that extracts of the anterior pituitary gland combat
and other techniques to determine the sequencing in the action of insulin. Frank Young found that adminis-
ribonuclease synthesized in the early 1960s. The pancreas tration of anterior pituitary extract causes a persistent
also produces another hormone, glucagon, a polypeptide diabetic condition. Cyril Long and Francis Lukens
containing 29 amino acids in a known sequence. discovered an antagonism between insulin and the secre-
The chemical structures of the steroid hormones tions of the adrenal cortex. These and other observations
produced by the adrenal glands and the sex organs are that hormones can act against each other have proved
related to cholesterol. They are all very similar and even useful in elucidating the endocrine control of the vital
small structural changes in their molecules produce functions and in medical and surgical treatments of certain
profound physiological eects. The steroid hormones conditions. However, it is now known that the substance
secreted by the adrenal cortex control carbohydrate and present in an endocrine gland does not necessarily take the
mineral metabolism; some are concerned with the forma- same form as that of the hormone in the blood.
tion of glucose from proteins and enable the body to Furthermore, hormones may themselves undergo meta-
withstand stresses such as intense heat or cold, injury and bolic changes, either in the blood or in the tissues, before
infection. The hormones of the adrenal cortex control they are able to cause reactions in the cells or in the enzymes
carbohydrate and mineral metabolism in the body. Similar they inuence. Thus, increased knowledge in endocrinol-
compounds, though with more complex structures, are ogy has created a situation in which the precise chemical
produced by the thyroid gland. Work on the sex hormones denition of a hormone has become a matter of some
was carried out from about 1926. In 19291930 Edward diculty.
Doisy in America, Guy Marrian in Britain and Adolf
Butenandt in Germany isolated from the urine of pregnant
women two ovarian hormones related to sterol. Butenandt
also isolated androsterone from male urine and in 1934 Vitamins
Leopold Ruzicka in Switzerland obtained this hormone
from cholesterol. These biologically active substances are Although dietary deciencies have long been recognized as
metabolites of the ovaries and testes, respectively. The the cause of certain diseases, the search for vitamins (a term
dierences in their molecular structures and physiological introduced by Casimir Funk) began about 1912. Gowland
actions are relatively slight; both are fat soluble. Hopkins observed that animals fed on a sucient quantity
The anterior pituitary gland inuences the rate of of puried foods ceased to grow unless a small amount of
secretion of the hormones of the adrenal cortex, the milk was added. This drew attention to the vitamin
gonads and thyroid gland. Its action is inuenced by nerve question, though for several years the real existence of
centres in the hypothalamus, the part of the brain these elusive dietary factors was disputed.
immediately above the pituitary gland. The secretion of In 1915 Elmer McCollum and M. Davis identied fat-
hormones and the functions they control are therefore soluble A and water-soluble B as essential accessory
coordinated by the nervous system. In some cases there is a dietary factors in rats. These were later named vitamins
feedback mechanism whereby the secretions of one A and B respectively. The latter was found to prevent beri-
endocrine gland stimulates or damps down secretions beri, while lack of vitamin A retarded growth and caused
from another. Thus a circle of action and reaction exists increased liability to infection of the respiratory system.
between some of the endocrine organs as the balance of The antiscorbutic vitamin C was later identied and in
hormone secretion is maintained. 1922 lack of fat-soluble vitamin D was recognized as the
The eects of injecting adrenaline are similar to those cause of rickets. About the same time vitamin E was
induced by stimulation of the sympathetic nervous system identied. In 1926 it was discovered that pellagra is a
and this led to the suggestion that the liberation of vitamin deciency disease. The vitamin in this case seemed
adrenaline at sympathetic nerve endings might transmit to accompany the anti beri-beri factor, but was dierent
the excitatory or inhibitory impulse to the eector cells of from it, in that it was much more stable to heat. In 1927 the
muscles or glands. Thus it seemed that certain hormones anti beri-beri factor was labelled vitamin B1 and the heat-
might control the functions of the central nervous system. stable factor became vitamin B2. This was at rst thought
It is now thought that a large number of chemicals can act to be a single compound, but was later found to be a
in this way, but only a few have been so far identied. The complex including riboavin, a yellow pigment with
rst of these, noradrenaline, was isolated by Hans van growth-promoting properties found in milk. In 1934
Euler in 1946. Others include acetylcholine, dopamine and pyridoxine, another component of the B2 complex was

6
 History of Biochemistry

identied. Nicotinic acid and nicotinamide (niacin and tion of the pigment present in the red cells. The nature of
niacin amide), the anti-pellagra factor, were identied in the active part of haemoglobin and the structure of the
1937. For several decades additional vitamins were porphyrin ring was investigated by Ernst Kuster in 1913.
discovered. Three are of particular importance for human Max Perutz and John Kendrew determined the complete
blood: vitamin K promotes the formation of prothrombin structure of the haemoglobin molecule using X-ray
in the liver, folic acid prevents anaemia and vitamin B12 spectrographic techniques between 1937 and 1959.
(cyanocobalamin) is the anti-pernicious anaemia factor. The white blood cells, or leucocytes, include a propor-
Each of the four fat-soluble vitamin groups, A, D, E and tion of phagocytes, cells that engulf and digest bacteria,
K, includes several related compounds with biological protecting the body from disease. In addition, 2025% of
activity. All contain one or more units of an isoprene the white cells, the lymphocytes, combine with antigens
structure in their molecules (C 5 CHC.(CH3) 5 CH). and remove them from the body, so controlling infections.
These vitamins are transported by lymph from the There are two types of lymphocytes, called B cells and T
intestines to the blood. Bile salts are required for their cells. The B cells produce chemical antibodies on activation
ecient absorption. They may be taken up as esters of by antigens and release them into the bloodstream. In the
palmitic acid or combined with a protein. Vitamins A, D 1970s the Japanese immunologist Susumu Tonegawa
and K are stored chiey in the liver; vitamin E is found in showed that about 1000 pieces of genetic material in the
body fat. The action of the fat-soluble vitamins is antibody-producing part of B lymphocytes can be shued
connected with certain enzymes. Some carotenes also and recombined in dierent sequences, enabling up to 1
show vitamin A activity. Only a and b carotenes and billion dierent types of antibodies to be formed each
tryptoxanthin are important in human metabolism and b specic to a dierent antigen.
carotene is the most active. Water-soluble vitamins include Serum albumin accounts for 55% of the total protein in
vitamin B1 (thiamin), B2 (riboavin), B3, B6 (pyridoxine), blood plasma. Its main function is to help maintain the
niacin, vitamin B12, folic acid, pantothenic acid and biotin. osmotic pressure between blood vessels and tissues.
In metabolic processes they act as coenzymes. Thus, Circulating blood tends to force blood out of the blood
vitamins B1, B2 and B6 become phosphates, biotin under- vessels and into the tissues, but the colloidal nature of
goes a change in structure and nicotinic, pantothenic and albumin and to a lesser extent of other blood proteins, the
folic acids form esters. Water-soluble vitamins are not globulins, keeps the blood within the blood vessels.
stored in the body to the same extent as fat-soluble ones Albumin also contains two materials necessary for the
and any excess is excreted in the urine. In addition to the control of clotting: antithrombin keeps the clotting enzyme
true vitamins there are other substances with vitamin thrombin from working unless needed and heparin
activity such as choline and p-amino benzoic acid, but these cofactor is necessary for the anticlotting action of heparin.
are fabricated in the body, as well as occurring in foods, The fate of cholesterol, another chemical substance found
and are not considered true vitamins. in the bloodstream, was investigated by Henri V. Goldstein
in 1972. He found that low-density lipoproteins, the
primary cholesterol-carrying particles, are withdrawn
from the bloodstream into the bodys cells by receptors
Blood Chemistry on the cells surface. Although dealing with complex
chemical substances and processes, studies on the blood
The dierence in colour between venous and arterial blood tend to be physiological and immunological, rather than
was known in ancient times, but that the same blood biochemical. They form part of medical research and it is in
changed colour as it circulated from veins to arteries such areas that the overlap between biochemistry, medical
through the lungs was discovered by William Harvey in the chemistry and physiology highlights the diculties of
1620s. The cellular structure of the blood was also rst demarcation and the precise denition of biochemistry as
observed in the seventeenth century. The red blood cells an independent discipline.
attracted most attention and in the nineteenth century it
was realized that they were the oxygen carriers. In 1827
Hans Fischer and his co-workers synthesized a large
number of porphyrins, the chemical components of the red Molecular Biology and the Nucleic Acids
cells, and showed how they transport oxygen in the blood.
As the only uid circulating through all the organs, the Since the 1950s progress in biochemistry has been led by
blood was thought to transport nutrients and remove molecular biology, a subject concerned with the ultimate
waste products, maintaining homeostasis in the body, physiological organization of living matter at the mole-
fundamental to Bernards theory of the internal environ- cular level. It is virtually impossible to distinguish between
ment. In 1862 Hoppe-Seyler observed the characteristic biochemistry and molecular biology, since both are
absorption spectrum of oxyhaemoglobin and in the same concerned with intermolecular transformations within
year William Stokes demonstrated the oxidationreduc- living cells. Since its inception in the late nineteenth

7
 History of Biochemistry

century biochemistry has sought to develop a molecular units. From this work a general theory of the diraction of
biology in stark contrast to the macrobiology of organs X-rays by helical structures was evolved. This would be
and tissues. Yet modern molecular biology seeks recogni- very important for determining the structures of the nucleic
tion as a separate discipline concerned with the molecular acids. During the same period another group of workers
basis of inheritance (genetics) and with protein synthesis in settled on the study of bacteria, believing that the
the cells. It has developed from and is related to bacteriophage was an ideal subject for the study of heredity
biochemistry, yet distinct from it. In 1950 W. T. Astbury because the transfer of hereditary material was not
identied molecular biology with a study of the forms of confused by other biological functions such as metabolism.
biological molecules, their development through higher The so-called phage group was indierent, even hostile, to
levels of organization and their relationship with genesis chemistry and perhaps due to this attitude it was only in
and function. The rst part of this denition is conforma- 1944 that it was realized that the materials under study
tional, the second is informational and for some years these were actually nucleic acids.
two aspects of molecular biology were studied indepen- The British and American researchers were proceeding
dently. independently of each other until James D. Watson, a post-
By the end of the nineteenth century biochemists had doctoral student from the phage group, came to work in
come to recognize that complex biological molecules held Cambridge. Watson was working on DNA (deoxyribonu-
the key to understanding vital processes in living cells. But, cleic acid), a compound rst isolated from pus cells in 1869.
as the methods of classical chemistry were inadequate to Its signicance as genetic material was recognized in 1944
determine the structures of large molecules such as when it was observed that bacterial DNA changed the
proteins, polysaccharides or nucleic acids, biochemists genetic material of other cells. In 1953 Francis Crick and
concentrated on the transformations of smaller molecules Watson proposed the double helix structure for DNA,
forming the components of these complex molecules. providing the conceptual framework for understanding
Molecular biologists, on the other hand, adopted physico- DNA replication and protein synthesis. It appeared that
chemical techniques such as X-ray diraction, developed the DNA molecule was composed of two helical phos-
in the 1920s at the Royal Institution in London by W. H. phate-sugar chains running in opposite directions and
Bragg who built up a school of crystallography including crosslinked at regular intervals by four organic bases
Astbury, J. D. Bernal and Kathleen Lonsdale. Using this always appearing in pairs (thymineadenine and cytosine
technique it was discovered that the structures of complex guanine). The links between the chains are formed by
molecules could be determined, but even this was simpler relatively weak hydrogen bonds and the separation of the
than the problems of explaining their functions in terms of chains leaves each one as a template for duplication using
their structures. Astbury applied X-ray crystallographic the small molecules brought to the cells by the blood.
techniques to bres and discovered that when a natural In 1955 S. Ochoa in Spain discovered an enzyme,
bre such as hair is stretched, its diraction pattern polynucleotide phosphorylase, capable of synthesizing
changes due to molecular rearrangements in its structure. ribonucleic acid (RNA). It was later found that this
Bernal turned to other living materials and with Dorothy enzyme degrades RNA in the cells, but under test-tube
Hodgkin and Perutz obtained diraction patterns for large conditions it runs its natural reaction in reverse. The
crystalline protein molecules. Perutz analysed the structure enzyme has enabled understanding of processes whereby
of the haemoglobin molecule and Kendrew made the rst hereditary information in genes is translated through RNA
three-dimensional analysis of the molecular structure of intermediaries into enzymes that determine the functions
myoglobin. ATP synthase, an enzyme responsible for and character of each cell. With discoveries such as these
synthesizing ATP, the universal energy carrier in living the informational molecular biologists could proceed
cells, was also examined. In the 1930s, however, further without further help from the chemical conformationists
progress was impossible due to the inadequacy of who were free to move on to other structural problems.
contemporary methods of analysis. Although the way in which molecular biology appar-
In America it was recognized that the helix was a ently developed seems to t this simple scenario, it is
common structural form for large molecules. As X-ray incomplete. The development of molecular biology in-
diraction analysis was unable to cope with such a cludes many more aspects of biochemical research like
structure Linus Pauling suggested that model building plant viruses such as Tobacco mosaic virus. These can be
would oer the only hope of solving these dicult readily extracted from plants and crystallized; they all
problems. For success, however, precise inter-atomic contain RNA. The mechanisms of nuclear division have
distances and angles were required. Pauling therefore also been important, as has the role of RNA in protein
applied X-ray diraction techniques to the analysis of synthesis. The contributions of these and other lines of
amino acids and small peptides in order to determine their biochemical research cannot be ignored in considering the
dimensions with minute accuracy. By the late 1940s he was origins of molecular biology. In 1978 the New York
able to use his results to construct models of large protein Academy of Arts and Science held a meeting to promote a
and polypeptide molecules by combining these smaller broader view focusing attention on the history of protein

8
 History of Biochemistry

research. Since then developments in molecular biology Florkin M and Stotz EH (19721979) Comprehensive Biochemistry, sect.
have involved such important advances as the rst VI, vols 3033. Amsterdam: Elsevier.
complete synthesis of a protein, the detailed mapping of Fruton JS (1990) Contrasts in Scientic Style: Research Groups in the
Chemical and Biochemical Sciences. Philadelphia: American Philoso-
the arrangement of atoms in certain enzymes, the elucida-
phical Society.
tion of intricate mechanisms of metabolic regulation and Holmes FL (1974) Claude Bernard and Animal Chemistry. Cambridge
the molecular action of hormones. These lines of investiga- MA: Harvard University Press.
tion are closely related to medical research and prominent Needham DM (1971) Machina Carnis; The Biochemistry of Muscular
recent researchers in molecular biology are largely drawn Contraction in its Historical Development. Cambridge: Cambridge
from this eld. They include immunologists Caesar Mill- University Press.
stein and Georges Kohler, geneticists Christiane Nusslein- Needham J (ed.) (1970) The Chemistry of Life. Cambridge: Cambridge
University Press.
Volhard and Eric Wieshaus, and Hugh Esmor Huxley,
Neuberger A and Deenen LM van (eds) (19831997) Comprehensive
who proposed the sliding lament theory of muscle Biochemistry, vols 3538, 40. Amsterdam: Elsevier.
contraction. Olby R (1994) The Path to the Double Helix: the Discovery of DNA
(Reprint of 1974 edn with additions). New York: Dover.
Teich M and Needham DM (1992) A Documentary History of
Further Reading Biochemistry 17701940. Leicester, UK: Leicester University Press.
Weatherall M and Kamminga H (1992) Dynamic Science: Biochemistry
Coley NG (1973) From Animal Chemistry to Biochemistry. Amersham, in Cambridge, 18981949. Cambridge: Wellcome Unit for the History
UK: Hulton. of Medicine.